1990 年 38 巻 1 号 p. 153-155
A simple kinetic method to assay L-asparaginase and L-asparagine with an ammonia gas-sensing electrode is described. The method is based upon the de-amination of L-asparagine by L-asparaginase from Escherichia coli, resulting in the production of ammonia. The initial rate (mV/min) of ammonia release is proportional to the activity of L-asparaginase and also to the concentration of L-asparagine in the presence of a large amount of the enzyme. Optimal temperature, buffer composition and pH for the assays are specified. L-Asparaginase was determined in the range of 0.4-1.6U in a 0.1 ml sample; the recovery was 98.1-103.8% for 16 determinations and σn was 1.59. L-Asparagine was determined in the concentration range of 1×10-4-1×10-3M with σn-1 1.92. The method was applied to the determination of 1-5×10-4M asparagine added to human serum with σn-1 1.96 for 5 determinations.
